Fig. 1

ACA mRNA is spatially localized to the posterior of polarized chemotaxing cells. a Representative maximum intensity projections of confocal fluorescent images of individual polarized WT cells depicting F-actin (green), DAPI (blue), and ACA mRNA (pink). The white arrows indicate the asymmetric distribution of the mRNA spots. Outline of the cells are shown in white dashed lines. Data are representative of at least 4 independent experiments. b-c Representative merged phase contrast and maximum intensity confocal fluorescent images depicting DAPI (blue) and ACA mRNA (pink), in WT (B) and ACA-YFP/aca ⁻ cells (C). The direction of migration of the cells (indicated by the dashed white lines) was determined by the position of the aggregation center towards which the cells were moving in these self-aggregating chemotaxis assays. The white arrows indicate the asymmetric distribution of the mRNA spots. Outline of the cells are shown in white dashed lines. Data are representative of at least 4 independent experiments. d-e Representative merged phase contrast and maximum intensity confocal fluorescent images depicting DAPI (blue) and cAR1 mRNA (pink) in WT (D) and cAR1-YFP/car1/3 ^−/− (E) cells. Data are representative of at least 4 independent experiments. f Each cell was manually bisected, defining anterior and posterior ROI based on both the orientation towards the aggregate center and the relative posterior enrichment of ACA-YFP in the cell. g Simulated and linear estimates of mRNA units across cells is plotted for ACA-YFP/aca ⁻ and cAR1-YFP/car1/3 ⁻ cells. The boxes show the 50% confidence region from the median (red line). The bars cover a region with 99% confidence level from the median. All data points beyond this confidence level are considered as outliers and are shown as red dots. The statistical significance was inferred by the t-test: * represents p < 0.05 and ** represents p < 0.01 (n_ACA = 33, n_cAR1 = 27)